CAJ | 학술논문

背景:有报道指出石墨碳纳米颗粒具有强大的吸附能力,只要尽可能将其控制在有效浓度范围内,石墨碳纳米颗粒会具有很好的细胞相容性及增敏效应.目的:了解石墨碳纳米颗粒的形态特征,观察石墨碳纳米颗粒其对体外培养细胞增殖与超微结构的影响.方法:取石墨碳纳米颗粒0.5 g,加100 mL三蒸水,振荡后微孔过滤,即为石墨碳纳米颗粒母液.取处于对数生长期的HepG2细胞、L02细胞、H17702细胞、3T3细胞,调整密度为5×10~(-7)L~(-1)接种于6孔板,0.5 mL/孔,加入含胎牛血清、青霉素、链霉素的RPMI-1640培养基1.5 mL,培养24 h后弃去旧液,设1～5号孔为实验组,分别加入质量浓度为25,10,7.5,5,0.25 mg/L石墨碳纳米颗粒培养液2.0 mL,设6号孔为空白对照组,不加石墨碳纳米颗粒溶液,继续培养24 h后终止培养.用原子力显微镜测量石墨碳纳米颗粒的粒径,电子显微镜观察石墨碳纳米颗粒的形态特征;用细胞计数板在光学显微镜下计数不同浓度石墨碳纳米颗粒对细胞数量的影响;透射电镜观察7.5 mg/L石墨碳纳米颗粒对细胞超微结构的影响.结果与结论:石墨碳纳米颗粒呈球形微粒,粒径约20 nm.与空白对照组比较,各浓度石墨碳纳米颗粒培养液组除HepG2细胞外,其余3种细胞数量基本都有所增加,其中7.5 mg/L石墨碳纳米颗粒培养液对L02细胞、H17702细胞、3T3细胞、HepG2细胞数量的影响最为显著(P<0.05).对7.5 mg/L石墨碳纳米颗粒作用后的细胞进行透射电镜观察,可见石墨碳纳米颗粒分布于细胞内部,如细胞质、细胞核、线粒体中,未见亚细胞结构受损及细胞凋亡坏死现象发生.证实石墨碳纳米颗粒对体外培养的细胞无损伤不良反应,且能够促进细胞生长增殖,其作用强度与质量浓度有关,7.5 mg/L为较佳质量浓度.
배경:유보도지출석묵탄납미과립구유강대적흡부능력,지요진가능장기공제재유효농도범위내,석묵탄납미과립회구유흔호적세포상용성급증민효응.목적:료해석묵탄납미과립적형태특정,관찰석묵탄납미과립기대체외배양세포증식여초미결구적영향.방법:취석묵탄납미과립0.5 g,가100 mL삼증수,진탕후미공과려,즉위석묵탄납미과립모액.취처우대수생장기적HepG2세포、L02세포、H17702세포、3T3세포,조정밀도위5×10~(-7)L~(-1)접충우6공판,0.5 mL/공,가입함태우혈청、청매소、련매소적RPMI-1640배양기1.5 mL,배양24 h후기거구액,설1～5호공위실험조,분별가입질량농도위25,10,7.5,5,0.25 mg/L석묵탄납미과립배양액2.0 mL,설6호공위공백대조조,불가석묵탄납미과립용액,계속배양24 h후종지배양.용원자력현미경측량석묵탄납미과립적립경,전자현미경관찰석묵탄납미과립적형태특정;용세포계수판재광학현미경하계수불동농도석묵탄납미과립대세포수량적영향;투사전경관찰7.5 mg/L석묵탄납미과립대세포초미결구적영향.결과여결론:석묵탄납미과립정구형미립,립경약20 nm.여공백대조조비교,각농도석묵탄납미과립배양액조제HepG2세포외,기여3충세포수량기본도유소증가,기중7.5 mg/L석묵탄납미과립배양액대L02세포、H17702세포、3T3세포、HepG2세포수량적영향최위현저(P<0.05).대7.5 mg/L석묵탄납미과립작용후적세포진행투사전경관찰,가견석묵탄납미과립분포우세포내부,여세포질、세포핵、선립체중,미견아세포결구수손급세포조망배사현상발생.증실석묵탄납미과립대체외배양적세포무손상불량반응,차능구촉진세포생장증식,기작용강도여질량농도유관,7.5 mg/L위교가질량농도.
BACKGROUND: Previous research has indicated that graphite carbon nanoparticles have a strong adsorbability. While, when the concentration is effectively controlled, graphite carbon nanoparticles also have well compatibility and sensitizing effect. OBJECTIVE: To observe the morphology of graphite carbon nanoparticles, and to investigate the effects of graphite carbon nanoparticles on cell proliferation and ultramicrostructure.METHODS: Graphite carbon nanoparticles (0.5 g) were put in 100 mL triple distilled water to obtain graphite carbon nanoparticle mother liquid after oscillation and microfiltration. HepG2 cells, L02 cells, HI7702 cells, and 3T3 cells in the logarithmic phase were adjusted to the concentration of 5×10~7/L and inoculated in 6-well culture plate with 0.5 mL per well. Thereafter, the cells were cultured with RPMI-1640 culture media (1.5 mL) containing fetal bovine serum, penicillin, and streptomycin. The original culture solution was removed after 24 hours. The 1-5 wells were considered as the experimental group, and 25, 10, 7.5, 5, 0.25 mg/Lgraphite carbon nanoparticles (2.0 mL) were respectively added into each well; while, the sixth well was considered as the blank control group without graphite carbon nanoparticles. The cells in the blank control group were cultured for 24 hours. Particle diameter was measured using atomic force microscopy; morphology was observed using electron microscope; effect of different concentrations of graphite carbon nanoparticles on cell number was detected using hemacytometry under optic microscope; the effect of 7.5 mg/L graphite carbon nanoparticles on ultramicrostructure was observed under transmission electron microscope. RESULTS AND CONCLUSION: Graphite carbon nanoparticles were around and 20 nm diameter. Compared with the blank control group, cell numbers except HepG2 cells were increased, especially the effect of 7.5 mg/L graphite carbon nanoparticles was greatest (P < 0.05). Transmission electron microscope indicated that graphite carbon nanoparticles were distributed into cells, including cytoplasm, nucleus, and mitochondrion; while, subcellular structure damage and cell apoptosis and necrosis were absent. Graphite carbon nanoparticles have no side effects on in vitro cultured cells and can promote cell proliferation, showing a dose-dependence correlation, especially the concentration of 7.5 mg/L.